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Objective:To analyze the clinical curative effect of protective sleep nursing on neonatal hyperbilirubinemia.Methods:Eight neonates with hyperbilirubinemia who were admitted from April 2019 to August 2019 were enrolled. They were divided into control group (40 cases) and observation group (40 cases) by random digits table method. Both groups were given routine nursing. On basis of control group, observation group was given protective sleep nursing. The clinical effect, sleep time, discomfort reactions and nursing satisfaction were compared between the two groups.Results:After nursing, the sleep time, crying time and bilirubin level were (18.67 ± 1.45) h/d, (0.82 ± 0.12) h/d, (191.58 ± 12.74) μmol/L in the observation group, and (17.63 ± 1.33) h/d, (1.05 ± 0.15) h/d, (202.42 ± 13.08) μmol/L in the control group, there were significant differences between the two groups ( t values were 3.343, 7.573, 3.755, P<0.05). The duration and regression time of jaundice were (5.26±1.24), (8.70±2.12) d in the observation group, and (7.14±1.18), (12.95±2.31) d in the control group, there were significant differences between the two groups ( t values were 6.946, 8.573, P<0.05). The good rate of sleep quality, incidence rates of vomiting, skin damage and needle falling out, and nursing satisfaction rate were 90.00%(36/40), 7.50%(3/40), 5.00%(2/40), 10.00%(4/40), 100.00%(40/40) in the observation group, and 72.50% (29/40), 27.50%(11/40), 22.50%(9/40), 32.50%(13/40), 87.50%(35/40) in the control group, there were significant differences between the two groups ( χ2 values were 4.021-6.050, P<0.05). Conclusions:The application of protective sleep nursing in treatment of neonatal hyperbilirubinemia can effectively prolong their sleep time, improve their sleep quality, which is conducive to improving their symptoms, reducing discomfort reactions.And satisfaction of their family members is relatively higher.
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Objective To investigate the effect of decitabine (DAC) on human acute myeloid leukemia (AML) cell line HL-60 and the regulating of natural killer (NK) cell activating receptor (NKG2D) ligands(NKG2DL), and to detect the molecular mechanism of JAK-STAT3-SOCS signaling pathway. Methods The effect of DAC on the proliferation of HL-60 was detected by using CCK-8 assay. The cell apoptosis was analyzed by using Annexin-V/PI double standard method. The expressions of receptor NKG2DL including MICA/B and ULBPs in HL-60 cells were detected by using flow cytometry (FCM). The killing activity of NK cells was analyzed by using carboxy fluorescein diacetate succinimidyl ester (CFSE). The expressions of JAK/STAT3 signaling pathway or molecules including STAT3, its upstream kinases JAK1, JAK2 and the negative regulator of STAT3,SOCS-1,SOCS-3 were examined by Western blot.Methylation level of the SOCS-1,SOCS-3 gene after the treatment of DAC was analyzed by using methylation-sensitive high resolution melting(MS-HRM). Results There was an obvious inhibitory effect of DAC on HL-60 cells. The cell viability of HL-60 treated with 0.2, 0.5, and 1.0 μmol/L DAC for 48 h was decreased by (25±11) %, (39±8) % and (50±7)%(P<0.01)respectively compared with those cells without DAC treatment.The incidence of apoptosis was (24.77±7.50) %, (27.10±4.48) % and (30.53±3.93) % after DAC treatment for 48h respectively, which were higher than that of untreated cells[(3.11±0.50)%](P<0.01).DAC induced a significant up-regulation of MICA/B, ULBP-1, ULBP-3 in HL-60 cells, and enhanced the sensitivity of HL-60 cells to NK cytotoxicity. Western blot results showed that a down-regulating expression of STAT3 and JAK1, JAK2 protein was detected, in addition to the phosphor-STAT3 and phosphor-JAKs in HL-60 cells after DAC treatment, but the expressions of SOCS-1 and SOCS-3 protein were increased. HRM results showed that DAC could inhibit the methylation of SOCS-3 gene. Conclusion DAC can inhibit the proliferation of HL-60 cells, upregulate the expression of NKG2DL and enhance the cytotoxicity of NK targeted to HL-60 cells, which might be related to the activity regulation of intracellular JAK-STAT3-SOCS signaling pathway.
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Objective To investigate the effect of decitabine (DAC) on human acute myeloid leukemia (AML) cell line HL-60 and the regulating of natural killer (NK) cell activating receptor (NKG2D) ligands(NKG2DL), and to detect the molecular mechanism of JAK-STAT3-SOCS signaling pathway. Methods The effect of DAC on the proliferation of HL-60 was detected by using CCK-8 assay. The cell apoptosis was analyzed by using Annexin-V/PI double standard method. The expressions of receptor NKG2DL including MICA/B and ULBPs in HL-60 cells were detected by using flow cytometry (FCM). The killing activity of NK cells was analyzed by using carboxy fluorescein diacetate succinimidyl ester (CFSE). The expressions of JAK/STAT3 signaling pathway or molecules including STAT3, its upstream kinases JAK1, JAK2 and the negative regulator of STAT3,SOCS-1,SOCS-3 were examined by Western blot.Methylation level of the SOCS-1,SOCS-3 gene after the treatment of DAC was analyzed by using methylation-sensitive high resolution melting(MS-HRM). Results There was an obvious inhibitory effect of DAC on HL-60 cells. The cell viability of HL-60 treated with 0.2, 0.5, and 1.0 μmol/L DAC for 48 h was decreased by (25±11) %, (39±8) % and (50±7)%(P<0.01)respectively compared with those cells without DAC treatment.The incidence of apoptosis was (24.77±7.50) %, (27.10±4.48) % and (30.53±3.93) % after DAC treatment for 48h respectively, which were higher than that of untreated cells[(3.11±0.50)%](P<0.01).DAC induced a significant up-regulation of MICA/B, ULBP-1, ULBP-3 in HL-60 cells, and enhanced the sensitivity of HL-60 cells to NK cytotoxicity. Western blot results showed that a down-regulating expression of STAT3 and JAK1, JAK2 protein was detected, in addition to the phosphor-STAT3 and phosphor-JAKs in HL-60 cells after DAC treatment, but the expressions of SOCS-1 and SOCS-3 protein were increased. HRM results showed that DAC could inhibit the methylation of SOCS-3 gene. Conclusion DAC can inhibit the proliferation of HL-60 cells, upregulate the expression of NKG2DL and enhance the cytotoxicity of NK targeted to HL-60 cells, which might be related to the activity regulation of intracellular JAK-STAT3-SOCS signaling pathway.
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Objective To study the cyclooxygenase (COX-2) ,nuclear factor-κB (NF-κB) and vascular endothelial growth factor (VEGF) expression and clinical significance in triple negative breast cancer .Methods From January 2010 to December 2014 ,breast cancer treatment in our hospital 100 patients for the study ,50 patients with triple negative breast cancer ,50 cases of non-triple neg-ative breast cancer was detected by immunohistochemistry ,100 cases of breast cancer were detected by immunohistochemistry in or-ganizations COX-2 ,NF-κB and VEGF expression of lymphatic vessel invasion and lymph vessel density and D2-40 mark detection , statistical analysis of relevant clinical and pathological information .Results COX-2 in triple negative and non-triple negative breast cancer were 76% ,70% ,the difference was not statistically significant (P>0 .05) ,VEGF triple negative breast cancer and non-triple negative breast lesions in cancerous lesions positive expression rates of 60% and 36% ,respectively ,which had significant difference (P<0 .05) .NF-κB in triple negative breast cancer lesions and non-triple negative breast lesions positive expression rate was 66%and 32% ,respectively ,which had significant difference (P< 0 .05) .Triple negative breast cancer NF-κB and VEGF ,COX-2 and VEGF expression was significantly positively related to breast cancer .Conclusion Radiation and chemotherapy is a major means of triple negative breast cancer postoperative treatment ,while inhibiting the NF-κB ,the expression of VEGF and COX-2 is expected to become the new target for treatment of triple negative breast cancer ,is worth exploring .
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<p><b>OBJECTIVE</b>To investigate the molecular mechanism of the growth inhibitory effect of matrine on K562 cells in JAK/STAT3 mediated signal pathway.</p><p><b>METHODS</b>Western blot analyses were performed to investigate the differential expression of JAK2, STAT3, phosphor-STAT3 (Tyr705 & Ser727) and phosphor-JAK2 proteins after matrine treatment in K562 cells with or without human recombinant interleukin 6 (IL-6) pretreatment. The expression of STAT3 response gene products such as Bcl-xL, Cyclin D1 and c-Myc, were investigated by Western blot and quantitative real time RT-PCR (qRT-PCR). Expression of IL-6, a potent upstream activating factor of JAK/STAT3 pathway, was analyzed by both real time qRT-PCR and ELISA.</p><p><b>RESUTLS</b>Western blot revealed that matrine treatment resulted in a strong down-regulation of phosphor-STAT3 both in Tyr705 and Ser727 sites or phosphor-JAK2 proteins expression without significant effects on the total STAT3 and JAK2 proteins. The expression of phosphor-Tyr705 STAT3 and phosphor-Ser727 STAT3 was decreased to 0.370 ± 0.172 in K562 cells treated with 0.5 mg/ml matrine for 48 h, respectively, from 0.690 ± 0.119 and 1.150 ± 0.263 in control cells, accompanied with a dramatical down-regulation of phosphor-JAK2 from 0.670 ± 0.137 to 0.049 ± 0.057 (P<0.05). In addition, it was found that the expression of Bcl-xL, Cyclin D1, c-Myc was decreased both at the transcription and protein level in K562 cells after matrine treatment. Matrine treatment resulted in a significant decrease in the expression level of IL-6 in K562 cells from (35.1 ± 1.93) to (10.74 ± 1.83) and (8.66 ± 1.24) pg/ml at the dose of 0.5 and 0.8 mg/ml, respectively (p<0.05). Matrine treatment could diminish the up-regulation of STAT3, JAK2, phosphor-STAT3 and phosphor-JAK2 protein following pretreatment with IL-6 in K562 cells.</p><p><b>CONCLUSION</b>Matrine exerts its anti-leukemia effect by interfering with the JAK2/STAT3 signaling pathway. The inhibition of IL-6 expression may play a pivotal role in the disruption of JAK/STAT pathway by matrine.</p>
Subject(s)
Humans , Alkaloids , Down-Regulation , Interleukin-6 , Janus Kinase 2 , K562 Cells , Quinolizines , STAT3 Transcription Factor , Signal Transduction , Up-RegulationABSTRACT
Objective To investigate the mechanism of matrine in inhibition of proliferation the proliferation of human chronic myeloid leukemia (CML) K562 cells via MEK-ERK signaling pathway. Methods Western blot was used to detect the expression of MEK1, ERK1/2, Shc and SHP2 (the signal effect molecules of MEK-ERK pathway) in K562 cells. The transcription and translation of bcr-abl and target protein (bcl-xL, Cyclin D1, c-myc and p27) were detected by RT-PCR and Western blot. Results Matrine was able to significantly inhibit the phosphorylation of MEK1, ERK1/2, Shc and SHP2 in K562 cells and suppress the protein and mRNA expression of bcr-abl. Moreover, the expressions of bcl-xL, Cyclin D1 and c-myc were down-regulated significantly, while the expression level of p27 (a negative regulator of cell cycle progression) was increased markedly after matrine treatment. Conclusions Suppression of the growth of human CML K562 cells is related to the inhibition of bcr-abl-mediated MEK-ERK pathway activity. The down-regulation of phosphorylated proteins or protein kinases activity in signaling pathways might be an important molecular mechanism in control the activity of MEK-ERK pathway.
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<p><b>OBJECTIVE</b>To probe matrine acting on natural killer cell (NK) activating receptor NKG2D ligands expression in CML cell line K562 and its underlying molecular mechanism.</p><p><b>METHODS</b>The expression of NKG2D ligands (major histocompatibility complex class I chain-related molecule A or B (MICA/B), UL16-binding proteins (ULBP) 1, 2, and 3 on K562 cells were analyzed before and after treated with matrine by FCM. The cytotoxic sensitivity of K562 to NK cell was detected by FCM after CFSE staining at different effect-to-target (E/T) cell ratios. The expression of signal transduction and transcriptional activator 3 (STAT3) protein as well as phosphorylated STAT3 (p-STAT3) were detected by western blot.</p><p><b>RESULTS</b>After treatment with matrine, ULBP1 and ULBP2 expression, especially ULBP2 on K562 cells significantly increased, with mean fluorescence intensity (MFI) increasing to 615 and 1614 by 220 and 615 in the untreated cells, respectively. There was no significant change for MICA or ULBP3 expression. Matrine enhanced the susceptibility of K562 cells to NK-mediated cell lysis. At the ratio of E/T with 5:1, the proportion of the killed K562 cells increased to 32.8%, 38.1% and 40.5%, respectively (after 0.2, 0.5 and 0.8 mg/ml matrine treatment) by 29.2% in the untreated cells. The phosphorylated STAT3 protein, but not STAT3 protein, was significantly inhibited by matrine treatment in K562 cells.</p><p><b>CONCLUSION</b>Matrine induced the expression of NKG2D ligands in K562cells and enhanced the cytotoxicity of NK cells against K562, which was closely related to the inhibition of STAT3 activity in K562 cell.</p>
Subject(s)
Humans , Alkaloids , Pharmacology , GPI-Linked Proteins , Allergy and Immunology , Intercellular Signaling Peptides and Proteins , Allergy and Immunology , K562 Cells , Quinolizines , Pharmacology , Signal Transduction , Up-RegulationABSTRACT
Objective To investigate the effects of N-acetylcysteine (NAC) on the expression of transforming growth factor-β1 (TGF-β1) in renal cortex of diabetic nephropathy rats.Methods A rat model of DN was established.The rats were randomly divided into control group,DN group and NAC group.After 8 weeks treatment,urinary albumin excretion rate (UAER) was determined.The expression of TGF-β1 in renal cortex was detected by immunohistochemistry and RT-PCR analysis.Pathomorphological changes of renal cortex were observed.Results (1)The levels of UA ER were significantly higher in DN group and NAC group [(1268.3±297.5) μg/24 h and (315.9-±86.8) μg/24 h] than in control group [(31.2±8.9) μg/24 h,q-29.85,16.76,both P＜0.01].The groups of DN and NAC versus group of control showed the increased levels of activity of TGF-β1 in renal cortex [immune-histochemistry index of glomerular mesangial area:7.35±1.17 and 3.87 ± 0.71 vs.1.95±0.34,q= 10.75,5.82,both P＜0.01];immune-histochemistry index of renal tubulointerstitium [21.21± 3.78 and 10.67±1.86 vs.3.62±0.79,q=15.20,11.36,both P＜0.01];the expression of mRNA in renal cortex[0.72±0.06 and 0.45±0.05 vs.0.23±0.04,q=9.13,7.45,both P＜0.01].The pathomorphological changes were significant in DN group and NAC group.(2)The NAC group versus DN group showed a decreased levels of UAER (q=8.17,P＜0.01),activity of TGF-β1 in renal cortex [immune-histochemistry index of glomerular mesangial area:q= 4.97,P＜0.01]immune-histochemistry index of renal tubulointerstitium (q = 6.86,P ＜ 0.01 );the expression of mRNA in renal cortex (q= 3.69,P＜0.05) and showed improvement of pathomorphology in renal cortex.(3) There was a significantly positive correlation between expression quantity of TGF-β1 mRNA in renal cortex and UAER level in NAC group(r= 0.749,P＜0.05).Conclusions The protective effects of NAC on the kidney of DN rats may be partly related with inhibition on the expression of TGF-β1.
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Objective To observe the effect of transfer factor on immunity in patients with varruca plana and the clinical effect of treatment. Methods Sixty patients with varruca plana were randomly divided into a treatment group (treated with transfer factor) and a control group (treated with routine therapy). Before and after the treatment, T-lymphocyte subgroups in peripheral blood of all the patients were determined by flow cytometry and serum levels of interleukin-10 (IL-10)and in-terferon-gamma (INF-γ)were detected by ELISA. The same detections were done to the twenty healthy volunteers as healthy controls. Results Effective rates in the treatment group and control group were 76.67 % and 43. 33 %, respectively (x2=5. 63,P<0.05). Decrease of CD3+ cells, CD4+ cells, CD4+/CD8+ ratio and increase of CD8+ cells were found in varruca plana group as com-pared with the healthy controls(P<0.01 ). After the treatment of transfer factor, increase of CD3+ cells ,CD4+ cells (P<0.05), CD4+/CD8+ ratio (P<0. 01)and decrease of CD8+ cells(P<0.01)were found in the treatment group as compared with those before treatment, while there were no significant difference in the control group before and after the treatment. Higher IL-10 and lower INF-γ in serum were found in varruca plana group as compared with the healthy controls (P<0.05). After the treat-ment of transfer factor, decrease of IL-10 and increase INF-γ in serum (P<0. 05)were found in the treatment group as compared with those before treatment, while there were no significant difference in control group before and after the treatment. Conclusion The results reveal that immunity is im-paired in patients with varruca plana. Transfer factor can enhance the immunity of the patients. Therefore, varruca plana patients treated with transfer factor receive better effects.
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Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC in different cells were measured by flow cytometry 48 h after transfection. Results: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells ([2.33±0.37] % and [5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 ([12.89±3.2] % for AdS-TRAIL and [9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells ([7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.
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Aim To investigate the effect of matrine on cytoskeletonof K562 cells. Methods Micropipette aspiration technique was adoptedto investigate the viscoelasticity of K562 cells, while the different ex-pression of cytoskeletal protein gene was analyzed by DNA microar-ray. Results In matrine-treated K562 cells, the viscoelastic propertiesKI, K2 and were decreased significantly from 726 ± 215 to 432 ±67,433 ±119 to 242±31, 72±38 to 50±15 respectively, and the geneexpression of prefoldin and ezrin was much stronger than that of controlcells. Conclusion The strueture and function can be changed in ma-trine-treated K562 cells.
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Objective:To study the anti-tumor effects of matrine on murine hepatocarcinoma cell line H 22 in vitro and in vivo .Methods:Cytotoxicity effect of matrine on cultured H 22 cells was determined by MTT assay in vitro .Apoptosis of cultured H 22 cells induced by matrine were determined with Annexiin V-FITC/PI affinity assay.Tumor-bearing BALB/C mice were used to observe the inhibitory effects of matrine on H 22 cells in vivo .The ultra micro-structured changes of H 22 cells were observed by transmission electron microscope in tumor-bearing BALB/C mice after treated with matrine.The expressions of Bcl-2 & Bax protein were detected by immunohistochemical technique and the staining densities of Bcl-2 & Bax protein were quantitated through computerized image processing.The data were analyzed with one-way ANOVA by means of SPSS 10.0.Results:Matrine could obviously inhibit the growth and induce the apoptosis of cultured H 22 cells.Matrine also had significant anti-tumor activity,on mice bearing H 22 hepatoma.The inhibitory rates were above 60.7% and 62.5% in higher and lower doses groups,respectively ( P